NMR of alpha-synuclein-polyamine complexes elucidates the mechanism and kinetics of induced aggregation

EMBO J. 2004 May 19;23(10):2039-46. doi: 10.1038/sj.emboj.7600211. Epub 2004 Apr 22.

Abstract

The aggregation of alpha-synuclein is characteristic of Parkinson's disease (PD) and other neurodegenerative synucleinopathies. The 140-aa protein is natively unstructured; thus, ligands binding to the monomeric form are of therapeutic interest. Biogenic polyamines promote the aggregation of alpha-synuclein and may constitute endogenous agents modulating the pathogenesis of PD. We characterized the complexes of natural and synthetic polyamines with alpha-synuclein by NMR and assigned the binding site to C-terminal residues 109-140. Dissociation constants were derived from chemical shift perturbations. Greater polyamine charge (+2 --> +5) correlated with increased affinity and enhancement of fibrillation, for which we propose a simple kinetic mechanism involving a dimeric nucleation center. According to the analysis, polyamines increase the extent of nucleation by approximately 10(4) and the rate of monomer addition approximately 40-fold. Significant secondary structure is not induced in monomeric alpha-synuclein by polyamines at 15 degrees C. Instead, NMR reveals changes in a region (aa 22-93) far removed from the polyamine binding site and presumed to adopt the beta-sheet conformation characteristic of fibrillar alpha-synuclein. We conclude that the C-terminal domain acts as a regulator of alpha-synuclein aggregation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Benzothiazoles
  • Binding Sites
  • Fluorescent Dyes / metabolism
  • Humans
  • Molecular Sequence Data
  • Molecular Structure
  • Nerve Tissue Proteins / chemistry*
  • Nerve Tissue Proteins / genetics
  • Nerve Tissue Proteins / metabolism*
  • Nuclear Magnetic Resonance, Biomolecular
  • Parkinson Disease / metabolism
  • Polyamines / chemistry*
  • Polyamines / metabolism*
  • Protein Structure, Secondary*
  • Synucleins
  • Thiazoles / metabolism
  • alpha-Synuclein

Substances

  • Benzothiazoles
  • Fluorescent Dyes
  • Nerve Tissue Proteins
  • Polyamines
  • SNCA protein, human
  • Synucleins
  • Thiazoles
  • alpha-Synuclein
  • thioflavin T